An efficient protocol is described for the rapid in vitro multiplication of an endangered medicinal plant, Caralluma diffusa, via enhanced axillary bud proliferation from nodal explants collected from young shoots of six-months-old plant. The physiological effects of growth regulators Benzylamino purine (BAP), Thidiazuron (TDZ), Kinetin (KIN), α- Naphthalene acetic acid (NAA) full strength of Murashige and Skoog’s (MS) medium on in vitro propagation were investigated. The highest number of shoots in initiation (4.16±0.30) and in subculture (17.33±0.33) and the maximum average shoot length (4.17±0.03cm) were recorded on MS medium supplemented with BAP (4.44 μM) and TDZ (0.90 μM) at pH 5.8. Rooting was best achieved on MS medium augmented with NAA (5.37 μM). The plantlets regenerated in vitro with well-developed shoot and roots were successfully established in pots containing decomposed coir waste, vermiculite and gardensoil (111ratio) and grown in a shade house with 93.83% survival rate.